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Posted 06/19/2015 04:55:04 EDT
Would you be my dad? :)
Posted 06/14/2015 10:44:00 EDT
How can I help? Love this idea and would like to contribute in some way? Perhaps offer suggestions in your next column. Thank.
Posted 06/10/2015 02:33:13 EDT
Thank YOU Ellen Goodman!
Posted 05/28/2015 10:24:21 EDT
Thank you....I see you've changed the headline from "20 is the New 60." My previous comment was in response to that. The current 1 may be more appropriate for what's written in this article.
Posted 05/28/2015 10:18:31 EDT
Not quite 60 (58), but my husband is, and this article doesn't come close to describing our current lifestyle. My parents & his (in their 80's & 90's respectively). But not ours. Nor any of the people more »
Posted 05/22/2015 10:23:53 EDT
just that this in person tonight. it's beautiful thank you to all the people who made this happen. everyone, worth checking out this memorial day weekend! more »
Posted 05/12/2015 10:57:16 EDT
I LOVE this! So much creativity combined with practical application combined with education about our future combined with research combine with..... Well, let's just say as an educator I'd give this more »
Posted 05/06/2015 10:22:05 EDT
what addicted teen? more »
Posted 04/03/2015 12:27:45 EDT
i'm w/ ms. conduct on this 1. i did my participate in seders, loved the first few, but then quite frankly, lost interest. and i study religion! if my side of the family (the 1 raised catholic, but no more »
Posted 04/01/2015 10:09:43 EDT
I read the late Mr Nyhan's letter again, and agreed w/ every sentiment. Then I wondered how many of the people he mentioned, intending to spotlight how far they'd come regardless of where they'd gone more »
Posted 03/16/2015 08:39:07 EDT
NO! I just had to say my piece. NO! NO! NO! more »
Posted 03/15/2015 10:24:37 EDT
I was raised around baseball and loved itâ¦.until the steroids became big theme & issue. It became so obvious, and seem to take forever to get exposed, and even now, it's only partially exposed. The more »
Posted 03/07/2015 08:47:24 EST
I've long believed in the benefits of napping - for those who know it helps. But it works best for me when I don't set a time limit on it. In fact, if I know I only or should have 30 minutes, I can't more »
Posted 02/19/2015 05:52:50 EST
Thanks. more »
Posted 02/19/2015 10:12:25 EST
My husband just Googled "Boston roof raking" and saw this article published Feb. 13, 2013 after the 2013 blizzard, "Why you should never rake a pitched roof". Here's the link for those interested: htt more »
Posted 01/03/2015 09:40:39 EST
Diane Patrick's description of how domestic abuse breaks one down is so spot on. Especially when she starts w/ "you don't see it comingâ¦." And I'll always appreciate that she identifies that it can more »
Posted 11/21/2014 08:41:19 EST
at the very least, some please get him to Best Friends Animal Society in Kanab Utah, where every dog finds a or is home! This wonderful dog should definitely find a home! more »
Posted 11/20/2014 09:25:13 EST
Who's the "we" in your question? more »
Posted 11/01/2014 10:50:59 EDT
John Dewey would be so proud! more »
Posted 10/26/2014 10:39:30 EDT
the other thing i don't get; every other endorsement the globe has made is in direct contrast to this one. where do you think baker stands on any of these? let's start w/ "yes on question 4; earned si more »
Posted 10/26/2014 10:05:10 EDT
my response to the person who said "stop with the gender card." no, i won't. gender is still a huge issue in this country, and i refuse to succumb to the pressure, subtle, passive-aggressive or overt more »
Posted 10/26/2014 09:48:28 EDT
furthermore, i feel like your endorsement is one i'd overhear at the "university club." not a place known for its diversity of opinions & members. more »
Posted 10/26/2014 09:45:10 EDT
sorry, but this is white male bias in clever disguise. while i've not been pleased w/ coakley's campaign strategy (again), i don't think she's the person you described here. her work as an a.g. has sh more »
Posted 09/15/2014 09:50:16 EDT
money, money, moneyâ¦.monay! more »
Posted 08/31/2014 05:25:48 EDT
a beaver? really? more »
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The Thinix software will be displayed in the Advantech booth # 4222 on the MICA-101 mobile clinical assistant (MCA) hardware devices and the Advantech PIT-1702 patient infotainment terminal.
"We are very excited to be given this opportunity to showcase our software and value our relationship with Advantech that is allowing us to bring this technology to market," stated Anthony Clark, Director of Business Development for Thinix.
The two Thinix products are the Thinix Touch for MCA Devices and Thinix Rx Touchscreen Healthcare Platform.
Thinix Touch for MCA Devices directly addresses the problem of using a touchscreen MCA device with both an operating system that is inherently not designed for finger navigation and with applications that are not always optimized for finger operation. Thinix Touch™ and leverages the power of Microsoft Windows and extend its usability immensely on the MCA hardware platform.
Thinix Rx Touchscreen Healthcare Platform is an extensible software platform used in the deployment of patient infotainment devices which are touchscreen bedside computing devices upon which a patient may access entertainment and education content in a simple and secure manner.
Thinix Rx and Thinix Touch are both currently offered for sale bundled with world class hardware options from leading manufacturer Advantech.
About Thinix:
Thinix is a leading developer of products and services for the healthcare, hospitality, professional audio-visual, agricultural, government, and education markets. Thinix technology includes secure computing technologies, kiosk software, touchscreen infotainment and entertainment systems, enterprise management platforms, and conference and meeting room systems. Thinix products are available through select resellers, integrators and distributors throughout North America.
# # #
Thinix is a leading developer of products and services for the healthcare, hospitality, professional audio-visual, agricultural, government, and education markets.


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text: Personalised Flush Slate House Sign by Nutmeg Home & Gifts £45$97.51€66.59$74.03 4 customer reviews(4) Made to order £18.50$40.08€27.37$30.43 delivery to United States Receive this item 5-10 working days from dispatch International delivery available house number or house name (write exactly as required i.e upper and lower case) (required) colourcolour (required)WhiteYellowMagentaVioletGreenPetrolSky Bluecolour (required)WhiteYellowMagentaVioletGreenPetrolSky Bluecolour (required)WhiteYellowMagentaVioletGreenPetrolSky Bluecolour (required)WhiteYellowMagentaVioletGreenPetrolSky Blue sizesize10cm x 10cm10cm x 15cm (+ £13)size10cm x 10cm10cm x 15cm (+ $28.17)size10cm x 10cm10cm x 15cm (+ €19.23)size10cm x 10cm10cm x 15cm (+ $21.38) quantity by Nutmeg Home & Gifts Location: Bedford, GB read seller's profile see seller's complete range more items from this seller Acrylic Door Number Sign £45$97.51€66.59$74.03 Personalised Slate House Number Sign £45$97.51€66.59$74.03 Personalised Wooden Door Number Sign £45$97.51€66.59$74.03 Pastel Patterned Butterflies Wall Stickers £14.50$31.42€21.45$23.85 Personalised Acrylic And Stainless House Sign £68$147.35€100.63$111.87 Personalised Stainless Steel House Sign £36$78.01€53.27$59.22 view all you may also like PRODUCT DETAILS CUSTOMER REVIEWS DELIVERY RETURNS can be personalised ask seller a question save for later Made by us in the UK, this slate house number or name is a simple, contemporary addition to your home. Whether you're looking for a number or a personalised house sign with the name of your home, our natural slate signs are high-quality and easy to put up. The gorgeous slate is embellished with vinyl text or numbers, available in seven colour options. Each sign is supplied with pre-drilled fixing holes, rawl plugs, screws and colour coded domed screw caps. All signs are made to order just for you, and take approximately 7 to 10 days to be dispatched. They're sent safely in protective packaging Available in two standard sizes: square or rectangular. We also offer a bespoke service for this product with a limited number of larger sizes available to enable further personalisation. With this service we also offer free artwork so that you can see how your sign will look prior to manufacture. If you are interested in this service please send us a message. made from: Natural untreated smooth slate 1.2cm thick with bevelled edges. Our slate signs are a natural product and each piece will differ slightly through natural markings and colour variations. To maintain this products appearance simply clean with a good quality glass cleaner. dimensions: Available in two sizes 10cm x 10cm 10cm x 15cm Other sizes are available please send an enquiry for more information. product code: 201732 4 customer reviews Wed 08 Jul 2015 Arrived very quickly and looks great! Thu 24 Oct 2013 No comment supplied. Sat 12 Oct 2013 the price of this item does seem expensive for the size of it, however it looks really nice - a great chucky bit of slate that look very stylish on my wall. It is also very well packaged so no risk of breakage by the post man. Wed 08 May 2013 No comment supplied. see all customer reviews hide some customer reviews read about our product rating system ratings guide trusted independent customer feedbackforum excellent bad good poor This product is dispatched to you by Nutmeg Home & Gifts. MAINLAND UK order now and receive this item by price Standard Delivery Monday 19 October £2.75$5.95€4.06$4.52 Tracked Two Day Delivery Friday 16 October £6.70$14.51€9.91$11.02 NON-MAINLAND UK estimated delivery price Highlands & Islands of Scotland 2-5 working daysfrom dispatch £2.75$5.95€4.06$4.52 Northern Ireland 2-5 working daysfrom dispatch £2.75$5.95€4.06$4.52 Other UK Islands 2-5 working daysfrom dispatch £2.75$5.95€4.06$4.52 INTERNATIONAL DELIVERY estimated delivery price Western Europe 2-5 working daysfrom dispatch £11.50$24.92€17.01$18.91 Rest of Europe 5-10 working daysfrom dispatch £11.50$24.92€17.01$18.91 USA & Canada 5-10 working daysfrom dispatch £18.50$40.08€27.37$30.43 Asia & Australasia 5-10 working daysfrom dispatch £19.50$42.25€28.85$32.08 Rest of the world 5-14 working daysfrom dispatch £19.50$42.25€28.85$32.08 You have 28 days, from receipt of cancellable goods, to notify the seller if you wish to cancel your order or exchange an item. Please note: goods that are personalised, bespoke or made-to-order to your specific requirements, perishable products and personal items sold with a hygiene seal (cosmetics, underwear) in instances where the seal is broken are non-refundable, unless faulty. Read more about our returns policy.Should you choose to return or exchange your order you will need to deliver the item(s) to the UK, where this seller is based. Nutmeg Home & Gifts's return policy


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I'll tell you all about it when I see you again
... to pick up Dad, and then we got Wing Stop for dinner since I was adamantly opposed to going to a restaurant in my plane attire. Plus Wing Stop is super yummy. After dinner I gave Mom and Dad their presents as well, and then Mom drove me back to my apartment. There, I found a super surprise - my cousin Mike and his wife Cathy gave me their London partition, which I've always loved! I'm so excited, but I can't decide where I want to put it. ...
Independence Day/Second Day In Washington
... on and on, some were special ones, like ones that had all been launched that showed the colours of the American flag! And ones that spelt out U-S-A! And then there was the finale. You can see it for yourself in the video on this page. All in all it had been the best day in Washington yet! Seeing other celebrations, other than Waitangi Day and ANZAC Day that marked a day of American history. It is also good to see how different countries celebrate, and how they ...
We may have bit off more than we could chew...
... FREE! The maps, however, $3! So we opted to take a picture of the maps on our phones and use that instead #frugaltraveltip.
The zoo exhausted all of us and took us the better part of the day. By the time we made our way to the Metro it was 4:30 and I knew we were all out of steam (and mama out of patience). We began our tripi back to the campground. I purcahsed a SmarTrip card thinking I was being so city Saavy. I swiped it getting on the bus where I was informed ...
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Washington DC
... been one of my favorite presidents. I went from here to the White House. The White House is obviously one of the most historical sites in the world. I was tired by the time I saw it from walking at least 15 miles today and even more tired because we had to walk all the way around the White House. It is a long walk and even longer nwo than months ago becausse in recent months, 3 people have jumped ...


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Horror, ghost stories and a little bit of fantasy/adventure is what I like to write. Have at you.
Favorite Authors and Inspiration: All the Horror greats! Algernon Blackwood, Ambrose Bierce, HP Lovecraft, William Hope Hodgson, MR James and of a more recent era, Richard Matheson and Joe R. Lansdale. And Stephen King is ok.
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Not yet done. My order is not getting filled.
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These authors contributed equally to this work.
Hepatobiliary Malignancies
Hypoxia inducible factor 2 alpha inhibits hepatocellular carcinoma growth through the transcription factor dimerization partner 3/ E2F transcription factor 1–dependent apoptotic pathway†
Article first published online: 7 FEB 2013
DOI: 10.1002/hep.26188
Copyright © 2012 American Association for the Study of Liver Diseases
Volume 57, Issue 3, pages 1088–1097, March 2013
Additional Information
How to Cite
Sun, H.-X., Xu, Y., Yang, X.-R., Wang, W.-M., Bai, H., Shi, R.-Y., Nayar, S. K., Devbhandari, R. P., He, Y.-z., Zhu, Q.-F., Sun, Y.-F., Hu, B., Khan, M., Anders, R. A. and Fan, J. (2013), Hypoxia inducible factor 2 alpha inhibits hepatocellular carcinoma growth through the transcription factor dimerization partner 3/ E2F transcription factor 1–dependent apoptotic pathway. Hepatology, 57: 1088–1097. doi: 10.1002/hep.26188
- †
Potential conflict of interest: Nothing to report.
Publication History
- Issue published online: 28 FEB 2013
- Article first published online: 7 FEB 2013
- Accepted manuscript online: 5 DEC 2012 12:00AM EST
- Manuscript Accepted: 12 OCT 2012
- Manuscript Revised: 11 OCT 2012
- Manuscript Received: 28 MAY 2012
Funded by
- National Institutes of Health. Grant Numbers: R01DK080736, R01DK081417
- Michael Rolfe Foundation for Pancreatic Cancer Research
- Major Program of the National Natural Science Foundation of China. Grant Number: no.: 81030038
- National Key Sci-Tech Project. Grant Numbers: 2012ZX10002011-002, 2013ZX10002011-004, 2012ZX0930100-007, 2012ZX10002013-005
- National Natural Science Foundation of China. Grant Numbers: nos.: 81071661, 81000927
- Shanghai New Project for Excellent Youth. Grant Number: no.: XYQ2011020
- Zhongshan Foundation for Youth. Grant Number: no.: 2012ZSQN-06
- Research Fund for the Doctoral Program of Higher Education of China. Grant Number: no.: 20100071120064
Hypoxia inducible factors (HIFs) are activated in many tumors and show either promoter or suppressor activity, depending on tumor cell biology and background. However, the role of HIF member HIF-2α remains unclear in hepatocellular carcinoma (HCC). Here, HIF-2α expression was measured in HCC and paired peritumoral tissues by quantitative real-time polymerase chain reaction, western blotting, and immunofluorescence assays, and the clinical significance was explored in 246 HCC patients. In cell culture, HIF-2α levels were up-regulated or down-regulated by use of expression or short hairpin RNA recombinant plasmid, respectively. Cells were analyzed by immunoblotting, chromatin immunoprecipitation coupled with microarray, coimmunoprecipitation, and immunohistochemical staining. In vivo tumor growth was analyzed in nude mice. We found that the average expression of HIF-2α was relatively low in HCC tissues, and the decreased level was associated with lower overall survival (P = 0.006). High HIF-2α expression in HCC cells induced higher levels of apoptosis and expression of proapoptotic proteins and inhibited cell and tumor growth. Furthermore, HIF-2α inhibited expression of the novel target gene, transcription factor dimerization partner 3 (TFDP3). TFDP3 protein was found to bind with E2F transcription factor 1 (E2F1) and inhibit its transcriptional activity through both p53-dependent and -independent pathways. Reintroduction of TFDP3 expression reversed HIF-2α-induced apoptosis. Conclusions: Data gathered from cell lines, tumorigenicity studies, and primary HCC samples demonstrate a negative role of HIF-2α in tumors, which is mediated by the TFDP3/E2F1 pathway. Our study provides evidence supporting a possible tumor-suppressor role for HIF-2α and has uncovered a mechanism that links HIF-2α to a fundamental biological regulator, E2F1. (HEPATOLOGY 2013)
Hepatocellular carcinoma (HCC) is one of the most common, aggressive malignancies and is the third leading cause of cancer-related deaths. The poor prognosis of HCC is mainly because of the rapid progression of this disease.1-3 Advances in treatment of this disease are likely to stem from a better understanding of its biology and behavior. As with most solid tumors, the hypoxic microenvironment exists in HCC as a result of an imbalance between oxygen supply and consumption in proliferating tumors.1 Significant evidences indicate that the hypoxia inducible factors (HIFs) play an important role in the pathogenesis and pathophysiology of HCC. Becasue HIF inhibitors are currently undergoing clinical evaluation as cancer therapeutics, a more-thorough understanding of the unique roles performed by HIFs in human tumor is warranted.4, 5
Cellular responses to low oxygen tension are mainly mediated by the activation of HIFs, which consist of a constitutively expressed subunit (aryl hydrocarbon nuclear translocator) and an oxygen-regulated subunit, mainly HIF-1α and HIF-2α. HIF-1α and HIF-2α promote adaptation of tumor cells to hypoxic stress by regulating the expression of genes involved in metabolism, angiogenesis, erythropoiesis, cell proliferation, and apoptosis.6-9 HIF-1α is found to be expressed at a higher level in dysplastic nodules and implicates a malignant transformation. We and others have previously shown that HIF-1α expression is significantly associated with an advanced stage and aggressive phenotype and indicates a poor prognosis in HCC.2, 3 An approximately 48% amino-acid sequence homology is shared by HIF-1α and HIF-2α10; however, much less is known about the HIF-2α isoform, and there have been inconsistent, and even conflicting, reports about its role in HCC. Some studies conclude that HIF-2α promotes hypoxic cell proliferation by enhancing c-Myc transcriptional activity,11 whereas others argue that knockdown of HIF-2α increases cell viability and growth by autophagy.1 It is known that HIF-2α correlates with vascular endothelial growth factor (VEGF) expression and indicates a poor prognosis in melanoma and non-small-cell lung cancers.12, 13 Other studies show that HIF-2α acts as a tumor suppressor in breast and brain cancer.14, 15
To date, there are limited published reports describing HIF-2α in HCC and the role of this protein is still controversial.1, 16 Here, we provide evidence for a possible tumor suppressor role for HIF-2α in HCC. Chromatin immunoprecipitation coupled with microarray (ChIP-on-chip) and additional assays indicate that transcription factor dimerization partner 3 (TFDP3) is a novel target of HIF-2α and inhibits E2F transcription factor 1 (E2F1) transcriptional activity through multiple pathways. This study has uncovered another mechanism connecting HIF-2α to E2F1 in the induction of apoptosis.
Materials and Methods
Patients and Specimens.
Tissues were collected and tissue microarray analyses were performed as previously described.17 Follow-up procedures are described in the Supporting Materials and Methods.
Cell Culture and Recombinant Plasmid DNA.
HCC cell lines used in this study, construction of short hairpin RNA (shRNA), and expression of HIF-2α recombinant plasmids are described in the Supporting Materials and Methods. The pcNDA3-TFDP3 plasmid was a gift from Yu Zhang's lab at Peking University. To select the stably transformed monoclonal cell lines, selective medium containing neomycin (pcDNA3.1 plasmids) or puromycin (shRNA plasmids) was used.
Real-Time Polymerase Chain Reaction Assay and Western Blotting.
RNA extraction, complementary DNA synthesis, quantitative real-time PCR (qRT-PCR) reactions, and western blotting were performed as previously described.18
Immunofluorescence Staining and Apoptosis Analysis.
Primary antibodies (Abs) used were as follows: anti-HIF-2α from Novus Biologicals (Littleton, CO); anti-Ki67 from Dako (Carpentaria, CA); and anti- caspase-3 from Cell Signaling Tecnology (Boston, MA).
To quantify the extent of apoptosis, cells were harvested with trypsin without ethylenediaminetetraacetic acid and visualized by Annexin V/fluorescein isothiocyanate/propidium iodide (PI) staining, according to the manufacturer's protocol (BD Biosciences, Rockville, MD).
ChIP-on-chip, ChIP-PCR, and Immunoprecipitation.
A complete protocol of ChIP-on-chip provided by NimbleGen Systems (Madison, WI) is included in the Supporting Materials and Methods. Predicted binding sites were confirmed using ChIP-PCR in the MHCC97L cell line. Immunoprecipitation (IP) was performed as previously described.19
Statistical Analysis.
Data were analyzed with SPSS 16.0 software (SPSS, Inc., Chicago, IL), as previously described.17 A P value <0.05 was considered statistically significant.
Expression Level of HIF-2α in HCC Patients and Its Clinical Significance.
To explore effects of HIF-2α in HCC, we first detected HIF-2α levels in tumor and peritumoral tissues. HIF-2α showed a mainly cytoplasmic staining in cancer cells, as previously indicated (Fig. 1A),16 and the average levels of HIF-2α on both protein and messenger RNA (mRNA) were significantly lower in tumor than peritumoral tissues (Figs. 1B,C). We further explored the clinical significance of HIF-2α expression in 246 HCC cases. The clinical and pathological characteristics of these patients were listed in Supporting Table 1. The percentage and intensity of positively staining cells were largely varying among cases, and tumors were classified as strong in 118 cases (47.9%) and moderately or weakly positive in 128 cases (52.1%) (Fig. 1D).13 Duplicate tumor samples showed a good level of agreement with respect to intensity and percentage of positively stained cells. Kaplan-Meier analysis revealed that patients with high HIF-2α expression levels in tissues had significantly longer overall survival (OS) rates than those with low expression (P = 0.006) (Fig. 1E).
HIF-2α Inhibits HCC Cell Growth Mainly through Apoptosis.
To establish whether HIF-2α plays a suppressor role in HCC tumor growth, human liver cancer cell lines MHCC97H and SMMC-7721 were studied. Both HIF-1α and HIF-2α expression were up regulated in the MHCC97H cell line, and the proliferation rate was lowered under hypoxia condition (Supporting Figs. 1A,B). To explore the role of HIF-2α in overcoming the effects of HIF-1α and other factors, we isolated high- or low-HIF-2α expression clones that had been transfected with expression vector pcDNA3.1-HIF-2α or shRNA inhibition vector pT2sh-HIF-2α, respectively (Fig. 2A,B). In these clones, HIF-1α levels did not vary significantly (Supporting Fig. 1C).
Initially, viability of transfected cells was investigated. High expression of HIF-2α in MHCC97H cells was associated with a slower growth rate, compared to the respective control, whereas cells transfected with anti-HIF-2α shRNA grew faster than control (Figure 2C). These differences were also observed in the SMMC-7721 cell line (Supporting Fig. 1D,E). HIF-2α also significantly slowed cell growth rate under hypoxic condition, regardless of whether with or without HIF-1α RNA interference (Supporting Fig. 1F,G). Next, we determined whether the altered growth rate is the result of an increased cell death rate. Annexin V/PI assays showed a dramatic apoptosis rate increase in the high-expression HIF-2α clone, compared to respective control (Fig. 2D). Confocal fluorescence microscopy revealed mitochondrial dysfunction and more cytochrome C released from HIF-2α-overexpressing cells (Fig. 2E). Multidomain proapoptosis B-cell lymphoma 2 (BCL-2) proteins Bax and Bak were expressed at higher levels in cells with high HIF-2α expression, relative to controls (Fig. 2F), which would facilitate the release of cytochrome C from mitochondria by forming pores in the outer mitochondrial membrane.20, 21 We also observed a higher amount of cleaved caspase-3, a key executer of apoptosis, in overexpressing HIF-2α cells (Fig. 2F), which would have been triggered by the cytochrome C release. The Bcl-2 homology domain 3 (BH3)-only proapoptotic proteins, including the Bcl-2-associated death promoter (Bad), p53 up-regulated modulator of apoptosis (Puma), and Bid were also found to be up-regulated in HIF-2α-overexpression cells (Fig. 2F). No significant differences were found in Ki67 staining between groups with different HIF-2α levels (Supporting Fig. 1H). Collectively, these data further confirmed that high levels of HIF-2α in HCC cells caused a cell-growth arrest through apoptosis.
Having established that high expression of HIF-2α in human HCC increases OS rates and induces HCC cell-growth arrest in vitro, we further tested the effect of HIF-2α in vivo. Clones with different HIF-2α expression were established (Supporting Fig. 2A). Tumors formed by cells with high HIF-2α level were smaller than mock controls, and tumors formed by cells with low HIF-2α level showed no significant changes in tumor weight, compared to respective controls (Fig. 3A,B). Cell proliferation and apoptosis measurements indicated that overexpression of HIF-2α induced more cleaved caspase-3 expression in tumor cells, compared to mock-treated cells (Fig. 3C). These in vivo data support HIF-2α's role of inducing apoptosis in HCC cells.
HIF-2α Augments Apoptosis by Inhibiting Expression of the Target Gene, TFDP3.
To uncover how HIF-2α affects apoptosis, we sought to identify possible targets using a ChIP-on-chip screen in MHCC97H cells (GEO accession no.: GSE37167). With a specific anti-HIF-2α Ab, we identified 470 target genes bound within their promoter regions, spanning 2.2 kilobases upstream and 500 base pairs downstream of the transcription start site, by HIF-2α. To validate screening results, we performed an independent ChIP experiment coupled with qRT-PCR (Fig. 4A).
The biological function and cellular component of HIF-2α-binding target genes were analyzed according to their ChIP-on-chip assay enrichment score. Genes with the highest enrichment scores were those that regulate cellular processes and are located in the nucleus (Supporting Fig. 3A,B). A substantial number of genes regulating apoptotic processes were found to be HIF-2α-bound genes. The top 10 genes were ranked according to their PeakFDR values and PeakScore values (Supporting Table 5). We observed a particularly strong suppression of TFDP3 mRNA (Fig. 4B) and protein levels in HIF-2α-overexpressing cells, compared to controls (Fig. 4C), and these effects were not noted when HIF-1α levels were varied (Supporting Fig. 3C,D).
The TFDP3 gene encodes a member of the dimerization partner (DP) family of transcription factors,22-24 which exerts a regulatory function by dimerization with the E2F protein. The TFDP3 protein is known to be highly expressed in HCC, but not in healthy liver tissues.23 We hypothesized that HIF-2α repression of TFDP3 expression could account for the high rate of apoptosis observed in the HIF-2α-overexpressing cell line. To test this idea, we overexpressed TFDP3 and/or HIF-2α in MHCC97H cells (Supporting Fig. 3E). The number of activated caspase-3-positive cells was decreased when TFDP3 expression was reestablished in HIF-2α-overexpressing cells (Fig. 4D,E). The expression of Bax and Bad was also down-regulated in these cells (Fig. 4F). TFDP1 and TFDP2 expression levels were tested to exclude the possibility of effects on E2F transcriptional activity (Fig. S3F). Collectively, these data show that HIF-2α significantly increases apoptosis by repressing the transcription of TFDP3, and the reintroduction of TFDP3 expression significantly reverses the effect.
HIF-2α Induces Apoptosis Through the TFDP3/E2F1 Pathway.
It is well known that DP proteins bind to E2F1, forming heterodimers that are essential for E2F high-affinity DNA binding and efficient transcriptional activity.24, 25 The TFDP3 gene was shown to colocalize with E2F1 by confocal microscopy (Fig. 5A). The physical interaction between E2F1 and TFDP3 was further analyzed by co-IP. In an E2F1 pull-down assay, TFDP3 bound directly to E2F1. Reciprocally, in an IP assay using anti-TFDP3 Ab, E2F1 was found to be bound (Fig. 5B). Knockdown of E2F1 expression by siRNA in HIF-2α-overexpressing cells increased their growth rate and decreased some proapoptotic gene expression (Supporting Fig. 3G-I). All these data suggest that E2F1 plays an important role in the induction of apoptosis by HIF-2α.
E2F1 has been reported to induce apoptosis through both p53-dependent and -independent pathways.26 Therefore, expression levels of key genes in these two pathways were determined. Cells were transiently transfected with both HIF-2α and TFDP3 recombinant expression plasmids, either in combination or individually. Although results showed that the mRNA level of TP53 did not change significantly (Figure 5C), its protein level greatly increased in cells transfected with HIF-2α and decreased to a normal level when cells expressed high HIF-2α and TFDP3 levels (Fig. 5D). The p14ARF gene has been reported to be regulated by E2F1 and interacts with murine double minute 2 (Mdm2), thus preventing p53 ubiquitination and subsequent degradation.27 Our results showed that p14ARF levels increased more than 2-fold in the clonal line with high HIF-2α expression and decreased nearly to normal level in cells that expressed both HIF-2α and TFDP3 (Fig. 5C). No significant expression changes were found for Mdm2 and p53 kinase ataxia telangiectasia mutated, which induce p53 phosphorylation28 (Fig. 5C). Expression of the apoptosis-stimulating protein of p53 1 (ASPP1), which directs p53 activity toward apoptosis,29 increased significantly in cells with a low TFDP3 expression level and decreased to a nearly normal level in cells that coexpressed HIF-2α and TFDP3 (Fig. 5C). These results suggest that HIF-2α-TFDP3-E2F1 increases apoptosis by increasing p14ARF and ASPP1 transcription, which further increases the p53 protein level.
Expression of some genes in the p53-independent apoptotic E2F1 pathway, such as the p53 homolog, p73,30 BID,25 and several caspases,32 were found to be significantly up-regulated in cells with high HIF-2α levels, whereas the restoration of TFDP3 expression significantly reversed effects on expression levels of those genes (Fig. 5C). Effects on apoptosis protease-activating factor 1,33 Puma, and the BH3-only protein encoding the gene Noxa,34 regulated by both E2F1 and p53, also showed a similar trend (Fig. 5C,D). Overall, both p53-dependent and -independent pathways play a role in HIF-2α-induced apoptosis. We further measured expression levels of TFDP3 and some representative genes of the E2F1 pathway in HCC specimens, which were with different HIF-2α protein levels determined by immunohistochemistry (IHC) staining. We found that patients with high HIF-2α protein expression had low expression levels of TFDP3, but high levels of proapoptotic genes (Figure 5E). Collectively, data from in vitro models and HCC patient samples have been implicated for the mechanism of HIF-2α-induced apoptosis (Figure 5F).
HIFs are detected in most solid tumors and are known to correlate with poor patient prognosis. The identification of genes, such as VEGF and CA9, in a HIF-activated pathway reveals a clear mechanism by which HIFs contribute to the survival and progression of cancer cells, and clinical therapeutic targeting of HIFs has emerged as a rational approach to treat solid tumors.35 Recently, studies have shown that HIF isoforms differentially regulate gene expression and sometimes have opposing functions during tumor progression. Therefore, experiments elaborating the precise roles of each subunit is needed.4, 10 HIF-1α has been widely explored and shown to regulate cell proliferation and angiogenesis and to be correlated with OS in HCC.2, 3, 36 However, studies on HIF-2α function in HCC are limited and these results sometimes are inconsistent.1, 16, 31 Therefore, we explored clinical significance by measuring gene expression in tumor samples from HCC patients as well as putative mechanism by altering cellular HIF-2α expression levels by overexpression or knock-down experiments.
We found that HIF-2α-induced HCC cell growth arrest and knockdown of HIF-2α expression increased cell viability in vitro. These effects have previously been observed in other cell lines under hypoxic conditions.1 Our clinical data further confirmed the effect. Through analysis of samples from hundreds of HCC patients, we found that HIF-2α expression in HCC tissues positively correlated with patient OS. Moreover, there was a significant negative correlation between tumor size and relative HIF-2α level (P < 0.001). Thus, our results suggest a possible tumor-suppressor role of HIF-2α in HCC.
With ChIP-on-chip screening and subsequent confirmatory assays, we showed that TFDP3 is a novel transcriptional target of HIF-2α, and its expression was significantly repressed by HIF-2α. Our findings link HIF's transcriptional activity to one of the least understood of the DP family, TFDP3. An important property that distinguishes TFDP3 from other DP members is its ability to inhibit E2F1-induced p53-mediated apoptosis.39 We found that the p53-independent pathway is also modulated by TFDP3. The TFDP3 protein is only expressed in HCC, but not in healthy liver tissues.23 Further exploring its role as a potential therapeutic target in HCC may uncover new applications.
The E2F family is best known for regulating cell cycle, proliferation, and apoptosis and is involved in signaling cascades induced by hypoxia.32 The level of E2F1 is up-regulated under hypoxic stimulation, and HIF-2α has been shown to increase E2F1 levels by c-Myc.11 Here, we showed that HIF-2α promotes E2F1 transcriptional activity and expression levels of E2F1-dependent target genes by inhibition of TFDP3 expression (Fig. 5F). This property of TFDP3 provides another connection between HIFs and E2F1.
HIF-2α exhibits a mixed nuclear/cytoplasmic localization pattern in HCC samples, and both the percentage and intensity vary greatly between patients.16, 41 HIF expression is known to be affected by many factors. For example, viral hepatitis infection and liver cirrhosis cause inflammatory cell infiltration and a hypoxic microenvironment, further modulating HIF expression patterns.37, 38 Therefore, the rate of viral hepatitis infection and the presence of liver cirrhosis need to be carefully considered when evaluating a study cohort's HIF expression profile. There were 82.9% hepatitis B virus–positive and 81.3% cirrhosis-positive patients involved in our study, whereas these rates were only 67.3% and 45.1%, respectively, in another HIF-2α study.16 In addition, there are other epidemiologic features, such as geographic regions, racial and ethnic groups, gender, and study methods, such as cell type, cultural method, and staining method used, that may also influence HIF-2α expression profile.
The role of HIF-2α in the development of cancer is not clear, and there is some discrepancy in the existing literature. One study showed that HIF-2α was correlated with angiogenesis and a poor outcome in HCC,16 whereas we and another group found that HIF-2α regulated autophagy and apoptosis, and high HIF-2α expression in HCC correlated with a good outcome.1 Diverse or complex roles of HIF-2α have also been found in non-small-cell lung cancer (NSCLC). HIF-2α appears to promote tumor epithelial-mesenchymal transition and is defined as a promoter of NSCLC.40 On the other hand, deletion of HIF-2α results in an increased NSCLC tumor burden by regulation of tumor-suppressor gene secretoglobin 3A1a expression.5
Reconciling these diverse roles is difficult. One possibility is that the tumor microenvironment markedly influences the active role of HIFs,1 as has been shown in astrocytoma. Loss of HIF-1α impairs astrocytoma growth subcutaneously, but it increases its proliferative and invasive properties in the brain.14 In HCC, hepatitis virus infection and liver cirrhosis are perhaps important factors, which significantly affect the tumor microenvironment.37, 38 Another possibility is that a tumor may employ a "stop and go" strategy to maintain its growth and survival, as suggested by Acker et al.15 When oxygen levels are limited, HIF-2α acts negatively on survival by inducing apoptosis, but, at the same time, promotes angiogenesis. Clearly, the mechanisms regulating HIF-2α function need to be further explored.
In conclusion, we have established that TFDP3 is a novel transcriptional target of HIF-2α and have uncovered another mechanism that links HIFs to a fundamental biologic regulator, E2F1. We have shown that HIF-2α induces HCC cell apoptosis by its effect on the TFDP3/E2F1 pathway and provided clinical evidences demonstrating a possible tumor-suppressor role of HIF-2α.
- 1Roles of hypoxia-inducible factor-1alpha (HIF-1alpha) versus HIF-2alpha in the survival of hepatocellular tumor spheroids. HEPATOLOGY 2010; 51: 2183–2192., , , , , , .
- 2
- 3The expression of hypoxia-inducible factor-1alpha in hepatitis B virus-related hepatocellular carcinoma: correlation with patients' prognosis and hepatitis B virus X protein. Dig Dis Sci 2008; 53: 3225–3233., , , , , , et al.
- 4HIF1alpha and HIF2alpha: sibling rivalry in hypoxic tumour growth and progression. Nat Rev Cancer 2012; 12: 9–22., , .
- 5HIF-2alpha deletion promotes Kras-driven lung tumor development. Proc Natl Acad Sci U S A 2010; 107: 14182–14187., , , , , , et al.
- 6HIF-1 and mechanisms of hypoxia sensing. Curr Opin Cell Biol 2001; 13: 167–171..
- 7The role of hypoxia-inducible factors in tumorigenesis. Cell Death Differ 2008; 15: 678–685., .
- 8HIF-1alpha versus HIF-2alpha in endothelial cells and vascular functions: is there a master in angiogenesis regulation? Cell Cycle 2009; 8: 3252–3253., .
- 9Endothelial deletion of hypoxia-inducible factor-2alpha (HIF-2alpha) alters vascular function and tumor angiogenesis. Blood 2009; 114: 469–477., , , , , , et al.
- 10A novel bHLH-PAS factor with close sequence similarity to hypoxia-inducible factor 1alpha regulates the VEGF expression and is potentially involved in lung and vascular development. Proc Natl Acad Sci U S A 1997; 94: 4273–4278., , , , , .
- 11HIF-2alpha promotes hypoxic cell proliferation by enhancing c-myc transcriptional activity. Cancer Cell 2007; 11: 335–347., , , , .
- 12Hypoxia-inducible factors 1alpha and 2alpha are related to vascular endothelial growth factor expression and a poorer prognosis in nodular malignant melanomas of the skin. Melanoma Res 2003; 13: 493–501., , , , , .
- 13Relation of hypoxia inducible factor 1 alpha and 2 alpha in operable non-small cell lung cancer to angiogenic/molecular profile of tumours and survival. Br J Cancer 2001; 85: 881–890., , , , , , et al.
- 14The hypoxic response of tumors is dependent on their microenvironment. Cancer Cell 2003; 4: 133–146., , , , , , et al.
- 15Genetic evidence for a tumor suppressor role of HIF-2alpha. Cancer Cell 2005; 8: 131–141., , , , , , et al.
- 16Prognostic significance of HIF-2alpha/EPAS1 expression in hepatocellular carcinoma. World J Gastroenterol 2007; 13: 3176–3182., , , , , .
- 17Intratumoral balance of regulatory and cytotoxic T cells is associated with prognosis of hepatocellular carcinoma after resection. J Clin Oncol 2007; 25: 2586–2593., , , , , , et al.
- 18Contextual synthetic lethality of cancer cell kill based on the tumor microenvironment. Cancer Res 2010; 70: 8045–8054., , , , , , et al.
- 19CD151 amplifies signaling by integrin alpha6beta1 to PI3K and induces the epithelial-mesenchymal transition in HCC cells. Gastroenterology 2011; 140: 1629–1641 e1615., , , , , , et al.
- 20Mitochondria and apoptosis. Eur J Biochem 1998; 252: 1–15., .
- 21Bax directly induces release of cytochrome c from isolated mitochondria. Proc Natl Acad Sci U S A 1998; 95: 4997–5002., , , , , .
- 22Human TFDP3, a novel DP protein, inhibits DNA binding and transactivation by E2F. J Biol Chem 2007; 282: 454–466., , , , , , et al.
- 23Large scale identification of human hepatocellular carcinoma-associated antigens by autoantibodies. J Immunol 2002; 169: 1102–1109., , , , , , et al.
- 24A functionally distinct member of the DP family of E2F subunits. Oncogene 2006; 25: 3212–3218., , , , .
- 25The role of the transcription factor DP in apoptosis. Apoptosis 2003; 8: 461–468., .
- 26E2F1 death pathways as targets for cancer therapy. J Cell Mol Med 2007; 11: 239–251..
- 27p14ARF links the tumour suppressors RB and p53. Nature 1998; 395: 124–125., , , , , , .
- 28E2F1 uses the ATM signaling pathway to induce p53 and Chk2 phosphorylation and apoptosis. Mol Cancer Res 2004; 2: 203–214., , , , , .
- 29ASPP1 and ASPP2 are new transcriptional targets of E2F. Cell Death Differ 2005; 12: 369–376., , , , , , .
- 30Role for the p53 homologue p73 in E2F-1-induced apoptosis. Nature 2000; 407: 645–648., , , , , , et al.
- 31Downregulating hypoxia-inducible factor-2α improves the efficacy of doxorubicin in the treatment of hepatocellular carcinoma.. Cancer Sci, 2012; 103: 528–534., , , , , , et al.
- 32Direct coupling of the cell cycle and cell death machinery by E2F. Nat Cell Biol 2002; 4: 859–864., , , , , , et al.
- 33Apaf-1 is a transcriptional target for E2F and p53. Nat Cell Biol 2001; 3: 552–558., , , , , , et al.
- 34Noxa, a BH3-only member of the Bcl-2 family and candidate mediator of p53-induced apoptosis. Science 2000; 288: 1053–1058., , , , , , et al.
- 35Biology of hypoxia-inducible factor-2alpha in development and disease. Cell Death Differ 2008; 15: 628–634., .
- 36Antisense hypoxia-inducible factor 1alpha gene therapy enhances the therapeutic efficacy of doxorubicin to combat hepatocellular carcinoma. Cancer Sci 2008; 99: 2055–2061., , , , , , et al.
- 37The tumor microenvironment in hepatocellular carcinoma (review). Int J Oncol 2012; 40: 1733–1747., , , , , , et al.
- 38Hypoxia-induced angiogenesis in human hepatocellular carcinoma. J Mol Med (Berl) 2002; 80: 703–714., , .
- 39TFDP3 inhibits E2F1-induced, p53-mediated apoptosis. Biochem Biophys Res Commun 2007; 361: 20–25., , , , , , et al.
- 40HIF2alpha cooperates with RAS to promote lung tumorigenesis in mice. J Clin Invest 2009; 119: 2160–2170., , , , , , et al.
- 41The expression and distribution of the hypoxia-inducible factors HIF-1alpha and HIF-2alpha in normal human tissues, cancers, and tumor-associated macrophages. Am J Pathol 2000; 157: 411–21., , , , , , .
Supporting Information
Additional Supporting Information may be found in the online version of this article.
|Supporting Information Figure 1. Expression of HIF-2α and its role in liver cancer cell. (A) The proliferation capability of MHCC97H cells cultured under normoxic or hypoxic (1% O2) conditions. (B) Expression of HIFs in MHCC97H and SMMC-7721 cells in different oxygen concentrations. (C) Expression of HIF-1α in MHCC97H clones with differing HIF-2α levels. (D) SMMC-7721 cells were transfected with pcDNA3.1-HIF-2α, pT2sh-HIF-2α, or the corresponding empty vector, then selected for G418 or puromycin resistance to isolate stably transfected clones. Left graph showed HIF-2α mRNA levels in representative monoclonal cells, error bars indicated standard deviation (n=3,* p <.01, two-tailed test). Right image: HIF-2α protein levels in representative monoclonal cells. (E) Cell proliferation as measured by CCK-8 in SMMC-7721 representative monoclonal cells. Results were expressed as the relative absorbance normalized to the one measured at zero time point. Error bars indicated standard deviation (n=6,*p <.01, two-tailed test). (F) The gene-silencing effect of siRNA targeting HIF-1α. Error bars indicate standard deviations. (G) Proliferation assay of cells with different HIF-1α or/and HIF-2α levels, cultured under 1% O2 condition. In the figure, si denotes siRNA 7146 targeting HIF-1α, and control cells were transfected with siRNA containing a scrambled sequence. Error bars indicate standard deviations. *p <0.05. (H) Cell proliferation measured by Ki67 in MHCC97H clone. Quantification of the relative percentage of Ki67 positive cells were shown in right image. Error bars indicated standard deviation (n=6, two-tailed test).
|Supporting Information Figure 2. HIF-2α expression in orthotropic xenografts. (A) HIF-2α mRNA expression was confirmed in tumors formed by MHCC97H monoclones. (B) CD31 staining in tumors.
|Supporting Information Figure 3. HIF-2α augments apoptosis by inhibiting the expression of TFDP3 Classification of HIF-2α-bound genes identified by ChIP-on-chip assay based on the biological function according to the enrichment score. It should be noted that some genes may be represented in more than one classified group.(B) HIF-2α target genes identified by ChIP-on-chip were classified based on cellular component according to the enrichment score. (C) HIF-1α levels in MHCC97H cells, which were transiently transfected by siRNA, pcDNA3-HIF-2α expression vectors, or control sequences. (D) TFDP3 levels were measured in MHCC97H cells with different HIF-1α levels. (E) The RNA level of TFDP3 and HIF-2α were tested in MHCC97H cells which was transiently transfected with pcDNA3.1- HIF-2α, pcDNA3.1-TFDP3 or mock plasmid. (F) The mRNA levels of other members of DP family were confirmed in the MHCC97H cells which were transiently transfected with HIF-2α (pcDNA3.1- HIF-2α), TFDP3 (pcDNA3.1-TFDP3) or empty vector (pcDNA3.1). (G) Gene-silencing efficiency of siRNA targeting E2F1. Error bars indicate standard deviations. (H) Proliferation of HIF-2α-overexpressing MHCC97H clones with different levels of E2F1. Error bars indicate standard deviations. *p<0.05. (I) The expression levels of apoptosis genes in HIF-2α-overexpressing MHCC97H cells with different levels of E2F1.
|Supporting Information
|Supporting Information Tables
Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.


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Upcoming EventsOct17Sat9:30 am Parks & People at Lake Mayer @ Lake Mayer ParkParks & People at Lake Mayer @ Lake Mayer ParkOct 17 @ 9:30 am – 12:00 pmJoin the Savannah Tree Foundation and community volunteers for the second of our Parks and People series! Beginning with stroll around the park with a certified arborist discussing the local trees, we will then provide some care[...]
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#YTGlobal Without Ever Leaving Home
We are happy to announce, that If you cannot participate in Youth Time Global Forum this year, but would like to support your friends and colleagues during the project presentations, or you are interested in the employment and education topics, you can still be a part of the event.
The Global Forum in Barcelona will be broadcast in real time from the 28th to 31st October, 2015. You will find the link on the web-site youth-time.org closer to date.
Moreover, you will have an opportunity to take part in the discussions during the Global Forum and ask speakers and experts your questions on Thursday, October 29, from 17:00 to 18:30 using our Facebook page and the website youth-time.eu.
Join us in Barcelona on-line!
Agenda For The Global Forum In Barcelona Unveiled
The Youth Time Global Forum in Barcelona will begin in about a month and will last three days. In the following paragraphs Youth Time offers a description of what participants can expect.
Day 1: Networking day
After the participants have arrived, registered and checked in at our venue, they will be taken to the city centre of Barcelona. Here they will take part in social activities, which have been specially prepared for our event to let the participants get acquainted with one another in a relaxed and enjoyable way. In the evening there will be an opening event with a buffet, a networking game, and a welcome party!
Day 2: Employment day
After breakfast we will open the Forum with speeches by prominent guests. Right after that the plenary sessions will begin, in which the day's topic – employment – will be discussed under the title: 'Challenging Youth Unemployment'. Among the keynote speakers are Chris Barnes, an entrepreneur who has founded a successful online advertising agency, as well as Felix Marquardt, an established writer for the Huffington Post as well as the founder of 'The Atlantic Dinners', where opinion makers from the fields of politics, business, and the arts gather to exchange ideas.
After inspiring speeches and discussions with our speakers, we will continue with the project presentations. These will be handled by young and ambitious participants who are eager to share their solutions to the employment issue. The projects will be evaluated by an expert panel as well as by the participants, who will have the opportunity to question the project presenters. At the end of the working day there will be an interesting workshop, to challenge the minds of everyone at the event.
Day 3: Education day
After breakfast we will go straight to the plenary sessions, in which the day's topic – education – will be discussed under the title: 'The Future of Education'. Among the keynote speakers you can expect Manish Dabas, chairman of the BRICS international Forum, as well as Silvia Crocitta, youth trainer and founder of the EuroDemos Youth Mobility NGO. After a break, we will continue with the project presentations, which this time will focus on the matter of education. At the end of the presentations, participants will vote for the winner of the event and will then conclude the program with another workshop, this one providing an overview.
Finally, in the evening everybody will have to dress up, before they will be taken to a special location for the official closing ceremony. Here the winners of the Youth Time Global Forum will be announced, and their victory will be celebrated with an incredible Spanish party!
How the Global Forum evolved from the Rhodes Youth Forum
Youth Time always strives for innovation and improvement, and this year the team has the challenging task of outdoing the experience of the last few years.
The program has been intensified in comparison with last year's event. First of all, the program is one day shorter, which leaves less time to decide who this year's winner will be. There will also be a block of time on the schedule for journalists to interview our speakers and guests, making speakers and journalists busier than ever. Finally there will be fewer working hours, and participants will be responsible for deciding the event winner.
More active role
In comparison with last year, when participants were divided according to their field of interest, this year everybody will see all the projects. This is simply the result of the fact that all participants will vote to select a winner and accordingly will have to take a look at all the projects. Also there will be an increased focus on networking, as there will be more time available for networking activities in the program. Finally, in order to facilitate networking, Youth Time has developed an app which should increase the interactivity among participants at the event.
To speak in the spirit of Daft Punk, get ready for a 'harder, faster, better' experience!
Youth Time Global Forum Teaser
The Youth Time Global Forum in Barcelona will be a unique event, which will gather together more than 100 participants from all over the world with the aim of coming to solutions for the most prevalent youth issues.
The topic of the annual event is The New Era of Employment and Education: Solutions for Change. The topic will be relevant to the location and the current realities on the ground there. Young people who are looking for inspiration and people who have innovative ideas on how to tackle these issues will be brought together at this event, to come to effective solutions for change that inevitably will contribute to a new era of employment and education.
The program will consist, first, of workshops and second, of project presentation meetings, in which participants and experts will evaluate the proposals the presenters have made. The program will also include roundtable discussions with prominent guests, and there will be time for social activities, in which participants will explore Barcelona and will have a chance to get to know each other
'Prepare for the Unknown Showing Itself as a Powerful Source for Growth'
We continue to acquaint you with a team of moderators and experts who will be working with us during YT Summer School. Meet Jaron Reisman. He is going to moderate 'Conflict Reconciliation' workshop. Jaron is absolutely outstanding trainer and entrepreneur from the Netherlands.


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Top Definition
to prepare oneself after an event for which one should have been prepared has taken place.
You always postpare yourself so you have not succeeded in your venture.
by uttam maharjan February 27, 2010
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Looks like the weather will be dry this week, Bull mountain should be dry enough to ride. If you like to night ride this is one for you. Thursday 3-2, meet in the parking lot around 7.
Route most likely will climb Bull, climb Bare Hare, descend Bare down to bottom(this is also a bail out area to get back to the parking lot),take the fs rd to the right and climb back up to Bull and then fly down to the parking lot. This route is right at 16 miles.
Everyones invited, bring a light with a long burn time and a backup.
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Thread: Night on Bull mountain 3-2
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Comment guidelines: Please be respectful to other people when making comments on this page. Bullying and abuse will not be tolerated and may result in your account being banned. Due to feedback from our user community, comments are now only available to registered users.
Can't Be Heaven (aka Forever Together) is a 2000 American family comedy film directed by Richard Friedman. The film stars newcomer Bryan Burke and Ralph Macchio. The film was loosely based on the 1968 movie Blackbeard's Ghost.
Register to update information, save favorites, post photos, news stories and comments.


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Proverbs 11:24 & 25 States
"The World of the generous gets larger and larger; The World of the of the stingy gets smaller and smaller. The One who blesses others is abundantly Blessed, those who help others are helped". (The Message Translation)
Download the Weekly Announcements ——>>>Download
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Pacific Inn of Redwood City
- Today
- Open 24 Hours
- Mon - Sun
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General Info
Property Location Located in Redwood City, Pacific Inn of Redwood City is minutes from Woodside Central Shopping Center and close to Malibu Grand Prix. This hotel is within close proximity of Menlo College and Bedwell Bayfront Park.
Rooms Make yourself at home in one of the 75 air-conditioned rooms featuring refrigerators and microwaves. Complimentary wireless Internet access keeps you connected, and cable programming is available for your entertainment. Bathrooms have jetted bathtubs and hair dryers. Conveniences include coffee/tea makers and irons/ironing boards.
Dining A complimentary hot/cold buffet breakfast is served daily.
Business, Other Amenities Featured amenities include a business center, a 24-hour front desk, and laundry facilities. Free self parking is available onsite.
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Under pressure: Is Marco's utility increasing water pressure to increase income?
City manager says "no," some residents remain skeptical
MARCO ISLAND - While Marco utility leaders turn up the water pressure, some Marco residents are turning up the pressure on officials saying the city is beefing up their water bills.
City Manager Steve Thompson and Public Works and Utility Director Rony Joel report problems of meeting minimum state water pressure requirements, particularly in the estates area and say they are turning up the pressure at the plants to meet those needs.
Water pressure has been an ongoing problem on the Island in recent years, but skepticism of increasing the pressure with the intent of increasing income is a relatively new concern.
Marco resident Tarik Ayasun hasn't yet returned phone calls for comment, however, in recent memos to Thompson, he indicated great concern over the issue.
"When I received my December water bill, I was in shock!" he wrote in the letter dated Friday morning.
Ayasun said he was out of town most of the billing period, and upon request of utility manager Jim Lang, Ayasun also confirmed with a plumber that he had no water leaks and confirmed with his landscaper that he had no irrigation malfunction.
"On Wednesday morning, 5 a.m., I was outside the house trying to get my paper. It was another shock. My sprinklers were like geysers," Ayasun continued.
He reported that his sprinklers were shooting water onto Dade Court, Inlet Drive and to the neighbor's house across the street.
Ayasun reported that after speaking with his sprinkler serviceman again, he learned that residents are contacting the sprinkler service company reporting an approximate 50 percent increase in water pressure causing burst sprinkler heads and other problems.
Rick Boynton, owner of Thompson Irrigation on Marco Island said he hasn't heard about those problems.
"If anything, we usually get calls that there is no pressure ... Too much is better than not enough," added the business owner.
Boynton recommends that if homeowners are experiencing too much pressure, which will add to water use and expense, that they can turn down their zone run times.
This reporter also attempted to contact other sprinkler companies on Island, including Collier Genie and S & S Sprinkler, but those contractors have not returned the phone calls as of print time.
Thompson, per information in his Weekly Update to council Thursday evening, is also aware of all of the issues.
"We do have properties close to falling below 20 psi (pounds per square inch) — the DEP (Department of Environmental Protection) requirement," Thompson reported.
Those requirements are needed for both fire suppression and use of second level or higher restrooms, he added.
"There has been a suggestion that the higher pressure is a deliberate attempt to generate higher consumption of water and higher revenues for the city," Thompson wrote Thursday.
The city recently increased water and sewer usage rates by 9.5 percent and has reported the anticipation of future annual increases compounding to more than 30 percent in the next few years to accommodate declining income and escalating costs. Those future increases have not yet been approved by council.
"I have been very clear in my direction to city staff that (increasing water pressure to increase income) is not and will not be a consideration," Thompson said.
There are projects that will need to be done to improve these water pressure issues, including a new water tank, new pipes and other improvements, he has reported.
Joel added that the utility committee reviewed and recommended approval of a four-year plan for these improvements.
"The increase (at the plant) from 75 psi to 78-80 psi is a short term solution. The current CIP (Capital Improvement Plan) and rate structure has a four year program for pipe upgrade of approximately $3.5 million in conjunction with the south water tank project that should be on line by mid 2011," Joel said.
Over a month, the increase is about 2 to 5 percent in water-use, he added.
"When the improvements are completed, we do anticipate that the reduced water pressure will reduce water use by approximately 5 to 8 percent.
"When certain areas of the Island were developed, water pipes were installed to meet the demand at that time and in the near future. As the Island has developed, the demand has increased. To meet that demand through the same pipe, requires us to increase the pressure," Joel said.
While it's not on the agenda, council members said they anticipate broaching the issue at their meeting scheduled 5:30 p.m. Tuesday in the Community Room, 51 Bald Eagle Drive.
Have an experience on the issue you can share, contact reporter Kelly Farrell, 213-5335, or email firstname.lastname@example.com.


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Look, deeply into the mirror
see, your reflection, and then beyond
we are all there, U, me and we, the whole cast
U and I, each, are the composite
the miraculous and tragic, juxtaposed
aggregating into what we perceive to be, we
We co-inhabit these storylines, divine
as we play and parlay, scribing & painting
upon parchment and canvas, creationally synched
When the veils, boundaries and walls
are dissolved and we embrace, the energy
we find ourselves, pulsing between polarities, fabricated
In the essence of magick*
beyond the constructed separations
i'll await, to meet you & celebrate
motionless pendulums.
{Molecular Rebelles.}
(Visited 18 times, 1 visits today)
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I'm a gypsy, seduced by words that capture emotions and emotions that transcend words. Writing affords me an opportunity to express my deepest feelings in the format I'm most comfortable. I write to unravel and understand myself, to shift my consciousness and also to touch a few others that may find meaning and connection, through the metaphors I've strung together while seeking to share, reflect and relate. I'm most impressed by the lyricists, poets, troubadours and scribes that write their own story, magicians all, weaving tales and reality with wicked efficacy.
Latest posts by Jim Fry (see all)
- Soul's Solstice. {poetry} - December 23, 2013
- eternally, she. {poetry} - December 15, 2013
- dissolving the molecules. {poetry} - August 29, 2013
- sandtime story. {poetry} - August 22, 2013
- exquisite entanglement. {poetry} - August 13, 2013


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BBB Accredited Business since 04/15/2011
Dr. Mark Jaffee, DDS
(303) 449-82992897 Valmont Rd Ste 200, Boulder, CO 80301-1341www.markjaffeedds.com
BBB Business Reviews may not be reproduced for sales or promotional purposes.
Request a Quote
BBB Accreditation
A BBB Accredited Business since 04/15/2011
BBB has determined that Dr. Mark Jaffee, DDS meets BBB accreditation standards, which include a commitment to make a good faith effort to resolve any consumer complaints. BBB Accredited Businesses pay a fee for accreditation review/monitoring and for support of BBB services to the public.
BBB accreditation does not mean that the business' products or services have been evaluated or endorsed by BBB, or that BBB has made a determination as to the business' product quality or competency in performing services.
Reason for Rating
BBB rating is based on 13 factors. Get the details about the factors considered.
Factors that raised Dr. Mark Jaffee, DDS' rating include:
- Length of time business has been operating.
- No complaints filed with BBB.
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Additional Informationtop
Licensing, Bonding or Registration
Many local municipalities, townships, and counties have registration, bonding and/or licensing requirements. The BBB encourages you to check with the appropriate agency to be certain any requirements are currently being met.
Permit and license requirements for regulated industries in the state of Colorado can be viewed at the following website: http://www.colorado.gov/oed/industry-license/
Type of Entity
Incorporated: December 1999, CO
Contact InformationPrincipal: Dr. Mark Jaffee (Owner)Customer Contact: Tracy Waisman
Business Category
Dentists, Dentistry - Cosmetic


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I was using nextgen but it seemed that it had some combatibility issues with my artisteer generaed theme so i switched to nextcellent. When i created my theme i put some boxes in a cell(created in photoshop) to put the thumbnails inside. However, when i put the code of my gallery the thumbnails don't allign correctly. They are left and i want somehow to control their position inside my box. I can see some css settings but i don't know exactly what to edit. Any help?
Here's a pic
Thanks in advance!


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Cover Girl Lash Blast Clump Crusher mascara
711 reviews
71% would repurchase
Package Quality: 3.8
Price: $$
Package Quality: 3.8
Price: $$
INGREDIENTSWater, Acrylates Copolymer, Glyceryl Stearate, Disteardimonium, Hectorite, Stearic Acid, Caranuba Wax, Propylene Glycol, Triethanolamine, Polyethylene, Lecithin, Propylene Carbonate, Polyvinyl Alcohol, Synthetic Wax, Oleic Acid, Alcohol Denat, Benzyl Alcohol, Panthenol, Phenoxyethanol, Methylparaben, Glycerin, Ethylparaben, Propylparaben, Ascorbyl Palmitate, Xanthan Gum, Tocopherol, Trisodium EDTA, Simethicone, Tocopheryl Acetate, Methylcellulose, Iron Oxides May Contain: Ultramarine, Titanium Dioxide
on 7/9/2015 3:32:00 PM
Age: 19-24
Skin: Acne-prone, Fair, Not Sure
Hair: Other, Other, Other
Eyes: Other
This is my favorite drugstore mascara! It separate and volumizes lashes well at a great price! I have a video that talks about this mascara along with other ones if you want to check it out.
on 7/3/2015 8:35:00 PM
Age: 18 & Under
Skin: Sensitive, Fair, Cool
Hair: Brunette, Other, Other
Eyes: Blue
AMAZING!!! THIS IS MY HG! I'm so bad at applying mascara but this wand/formula makes it so easy and I always end up with flawless lashes! I have tried well over 30 different mascaras over the past couple years and this one is the best by far! I'v repurchased it so many times and I will continue to repurchase until its discontinued.
on 7/1/2015 10:30:00 AM
Age: 25-29
Skin: Normal, Fair-Medium, Not Sure
Hair: Blond, Other, Other
Eyes: Blue
This is my hg mascara!!!! Even after applying 3 coats, my eyelashes are clump free and perfect! I store old bottles of this to use the wand on other mascaras because the wand is so great. The formula is also amazing. It lasts all day and doesn't even move if I accidentally rub my eyes! I defiantly reccomend this mascara to anyone who loves long thick natural looking lashes for a cheap price!!
on 6/25/2015 11:08:00 AM
Age: 25-29
Skin: Oily, Fair, Warm
Hair: Brunette, Wavy, Medium
Eyes: Brown
Clump Crusher is a good basic, everyday mascara. It does a good job at separating the lashes and, true to its name, does not clump. I find that it stays on well and does not flake. If you're looking for a really dramatic look, though, this may not be adequate, as I don't find it does much in the volume department.
My tube smells a bit like paint but it's not too strong.
The wand on the Clump Crusher mascara is curved, which for me is kind of an awkward shape for my eye. I think I would find a straight wand easier to use. That's really my only gripe with it.
on 6/20/2015 9:50:00 PM
Age: 19-24
Skin: Sensitive, Fair, Not Sure
Hair: Blond, Other, Other
Eyes: Other
I find this mascara to be really lengthening, a natural looking mascara
on 6/19/2015 11:17:00 PM
Age: 36-43
Skin: Combination, Fair-Medium, Not Sure
Hair: Brown, Other, Other
Eyes: Brown
I really like this one! It truly does not clump, I mean, I'd have to work really really hard for it to clump on me, and I love that. I like the wand, it's rubber with short bristles, it doesn't hurt me and it separates the lashes very well. It's also curved, which I think helps to curl the lashes, it's easy to get very close to the lash line, and it's very good for the bottom lashes too.
I got the non-waterproof version and I think it holds up really well, it doesn't flake or smudge on me all day. And, it's easy to remove at the end of the day. Yay!
It's not extra volumizing or lengthening, but it gives me enough oomph and separation for everyday wear.
However, I don't think this mascara is for everyone. I do have full and naturally curvy lashes. So, if you have short or thin lashes and are looking for that "wow" transforming mascara, I think you might be disappointed.
on 6/19/2015 8:58:00 PM
Age: 18 & Under
Skin: Oily, Medium, Not Sure
Hair: Brown, Other, Other
Eyes: Brown
I LOVE THIS PRODUCT SO FREAKING MUCH OMFG!! it does exactly what it says it does, my lashes have no clumps at all. it's perfect and it is the only mascara i will ever use!
on 6/18/2015 9:25:00 PM
Age: 19-24
Skin: Oily, Fair-Medium, Cool
Hair: Other, Wavy, Coarse
Eyes: Brown
I hated this. Oh my god, I hated this so much.
I bought this because so many people raved about it. I don't know why.
I took it home, opened it up, applied it to clean eyelashes, and watched in terror as it clumped all my eyelashes together in spikes. I thought it might need to be mixed or something, so before I tried it the next time, I swirled the wand around in the bottle a bit. Nope. Clumped together so badly.
This wasn't an average mascara, this wasn't even an acceptable mascara. This was a terrible, unusable mascara. How it's gotten so much positive attention is beyond me. I ended up returning it.
on 6/9/2015 6:08:00 PM
Age: 19-24
Skin: Normal, Fair-Medium, Cool
Hair: Blond, Wavy, Medium
Eyes: Blue
If you're looking for a dramatic, eye enhancing mascara, this is NOT it. It's not horrible though. Does what it says, will not clump and I'd grab this if I was completely out of mascara. So/so product.
on 5/14/2015 10:13:00 PM
Age: 25-29
Skin: Dry, Fair, Cool
Hair: Brown, Curly, Fine
Eyes: Green
There have been several mascaras over the years with this type of brush and formula, but this one is in a Lash Blast tube so it's more exciting I suppose. I find this type of brush painful and always manage to poke myself hard in the eye every day I use it. I don't have very long nor curly eyelashes, so while this does create clean lashes it does not give me much volume nor curl. It isn't billed for that, so I am only down voting it because the wand is so darn painful! This may be due to the fact I wear glasses and cannot see perfectly when I apply mascara. I also get a good bit of smudging on my eyelid during application that messes up my eyeliner. I would not repurchase.
Popular Mascaras
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My Mascara Collection! The Good vs. The Bad by TheKatalicious17


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Select Local Merchants
Rich and Charlie's Italian RestaurantRead MoreFor spaghetti and meatballs that tug at the heartstrings, Rich and Charlie's Italian Restaurant creates a warm atmosphere perfect for two. Rich and Charlie's Italian Restaurant worries about making delicious food, not counting calories. Let the kids come too! Little ones love the food and atmosphere at Rich and Charlie's Italian Restaurant just as much as their parents do. Sit outside at Rich and Charlie's Italian Restaurant and soak up the sun on those nice summer days. Enjoy the vibe here with a business casual dress code. For the tastes of Rich and Charlie's Italian Restaurant from the comfort of your next party, the restaurant also offers catering services. If time is of the essence, Rich and Charlie's Italian Restaurant's take-out option may be a better fit. Rich and Charlie's Italian Restaurant is a local eatery that serves up free parking. Store your bike at one of the many racks outside of Rich and Charlie's Italian Restaurant. Your bill at Rich and Charlie's Italian Restaurant will typically run less than $30 per person, so bring the whole gang!4487 Lemay Ferry RdSaint Louis, MO US
China WokRead MoreFresh and flavorful Chinese favorites flood the menu at Saint Louis' China Wok. China Wok is a popular restaurant for those who prefer to eat low-fat and healthy fare. Carry-out is also available for those who prefer to enjoy China Wok's cooking from the comfort of their own home. Take the comfort of your own home and add great grub from China Wok to create the perfect night. Complimentary parking is provided in the lot next to China Wok. Prices tend towards the moderate side, with the average tab at China Wok running under $30 per person.4319 Telegraph RdSaint Louis, MO US
SubwayRead MoreFor tasty sandwiches with endless toppings, visit Subway. The food at Subway is high-calorie, high-flavor, and highly-rated. If you need to get somewhere fast, the restaurant also serves up grub to go. Subway prides itself in its delicious catering. Take advantage of the quick and easy parking near Subway. At Subway, diners can make use of the safe bike rack. Prices are downright affordable at Subway, with most items well under the $15 mark. Chow down on breakfast, lunch, or dinner fare at Subway — they're open for all three meals.2 Professional Park DrMaryville, IL US
Putter's Bar and GrillRead MorePutter's in Maryville is a well-known restaurant, offering a familiar vibe with appetizing burgers. It's a local favorite for customers looking for good food. If you just feel like stopping by for a beverage, the restaurant has helpful bar staff (and a good selection behind the bar). You won't walk away from Putter's disappointed.7700 Stonebridge Golf DrMaryville, IL US
T.G.I. Friday'sRead MoreT.G.I. Friday's proves that quick and easy all-American grub has its place in the ranks, too, and rave reviews flood through the front door. The menu doesn't include any low-fat items, so set aside some extra calories for your visit. T.G.I. Friday's also operates a bar, so a round of drinks with dinner is not out of the question. Take the kids along too — T.G.I. Friday's is a great spot for families with food that even little ones will love. Don't stay inside on a beautiful day! Come sit on the patio at T.G.I. Friday's and order great food. Don't go off the grid! With the free wifi at T.G.I. Friday's, you can surf the web and get some work done. Be prepared to raise your voice, though — the restaurant can get noisy. No need to gussy up for a trip to T.G.I. Friday's, where patrons dress for comfort and fun. You can also grab your food to go. Can't get enough of T.G.I. Friday's' tasty dishes? They also offer a catering service for parties and events. T.G.I. Friday's' diners will appreciate the free parking in a lot next door. Cyclists will love the spacious bike racks outside of T.G.I. Friday's. T.G.I. Friday's is creating dishes any foodie will love at around $30. T.G.I. Friday's has three square meals a day on the menu, so swing by for breakfast, lunch, or dinner.5262 S Lindbergh BlvdSaint Louis, MO US
Dulany's Grille and PubRead MoreFor bar nibbles and pub food par excellence, Dulany's Grille and Pub is a top pick. Low-fat is one factor and amazing taste is another. Come see which wins out at Dulany's Grille and Pub. With Dulany's Grille and Pub's wide selection of refreshments available, you can tap into the drink menu early in the evening. Save money on a sitter — kids are welcome to join the table at Dulany's Grille and Pub. Fridays are for fun! Come check out the vibe at Dulany's Grille and Pub. During the summer months, don't miss out on Dulany's Grille and Pub's outdoor patio seating. Dulany's Grille and Pub is a fine restaurant for those with large and small parties. Hop online in no time using Dulany's Grille and Pub's free wifi. Fancy-schmancy attire is not required; in fact, guests are told to keep things casual. Dining out isn't your only option here — pickup is available, too. Catering from Dulany's Grille and Pub will take your party to the next level. The lot adjacent to Dulany's Grille and Pub provides free parking for diners. Treating yourself doesn't mean breaking the bank, come taste the great dishes Dulany's Grille and Pub has to offer. Patrons can choose to charge their bill, as Dulany's Grille and Pub welcomes the use of most major credit cards.9940 Kennerly RdSaint Louis, MO US


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Graduate Admissions: Online Information Sessions
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I can't find a definitive answer anywhere- can I use something like Rival cranks with the 34t small ring on my mountain bike? I am under the impression that they might be too narrow and rub on the chainstays. otherwise, there's only one ring and no derailleurs to worry about.
Edit: my frame has a standard 68mm threaded BB.
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Poyourow Family in Minnesota from Ukraine 1920-?
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- Hello,
I am in search of the family members listed below. I have no further
information. I do know that some information on Social Security
documents and death certificates are wrong. They list the wrong
states etc. There are no graves to visit, they cremated them. There
is ashes out at sea but thats it.
What do I do? Thank you in advance
ESP of MD
Searching for my brick wall, Jacob Poyourow's parents. Jacob b2/14/1889 d10/1980 in Florida married Anna Schwartz b2/16/1897 d3/1975 in Florida, where and when married unknown. What I do know is Jacob entered into the U.S. through Minnesota. They had Lester Jacob Poyourow b8/18/1923 in Minnesota and d7/6/1983 in Florida. They also had Marvin Poyourow birth unknown date or place. There are 27 living Poyourow's in the USA as of today. Only 5 are related to Jacob unless I can prove who his parents were.
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from the cassette "The Spilling Past" on Bathetic Records.
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